ZhenQi FuZheng Granules Inhibits The Growth Of Colorectal Tumors Based On The Regulation Of Intestinal Flora

Colorectal cancer（CRC）is the fourth most common cancer in the world,and its high morbidity and mortality affect human life seriously.Conventional treatments strategies are surgery,chemotherapy,radiotherapy,radiochemotherapy and immunotherapy.However,due to the high cost of surgery,poor prognosis,recurrence,low response rate to immunotherapy,and decreased quality of life caused by subsequent radiotherapy and chemotherapy,effective treatment of cancer is not feasible.Therefore,there is an urgent need for alternative therapeutic agents for colorectal cancer with fewer side effects and lower cost.Zhenqi Fuzheng granules（ZQFZ）are mainly composed of Astragalus and Ligustrum lucidum,which have immune system regulation functions and potential anti-tumor biological activities.ZQFZ has been shown in previous studies to have ameliorating effects on cyclophosphamide-induced myelosuppression in hematopoietic system,showing its immunomodulatory activity again.The purpose of this experimental study was to investigate the effects of ZQFZ on CRC tumors through in vivo experiments in colorectal cancer mice and in vitro experiments with LPS-stimulated macrophages.In this study,ZQFZ observably reduced the phosphorylation of the protein belonging to the nuclear factor kappa-B inhibitor kinase（IKK）/nuclear factor kappa-B（NF-κB）signaling pathway in RAW264.7 cells induced by lipopolysaccharide（LPS）in vitro,exhibited anti-inflammatory activity of ZQFZ.In vivo experiments,we selected the B6/JGpt-Apc^{em1Cin（Min C）}/Gpt(Apc^Min/+)colorectal cancer mouse model with Apc gene mutation,and performed ZQFZ gavage treatment for 56 consecutive days.After administration,ZQFZ can slow down the formation and progression of intestinal tumors directly,reduced the size and number of tumors,alleviated the pathological changes caused by cancer in the liver,lung and colorectum and delayed the progression of CRC.ZQFZ altered the composition of gut microbiota in Apc^Min/+mice,especially the abundance of some bacteria that are related to the immune response closely,such as Bacteroidetes,Firmicutes,Actinobacteria,and Verrucomicrobia,suggesting that ZQFZ may mediate immunomodulatory effects through the intestinal flora.Based on this conclusion,subsequent studies on immune function found that ZQFZ significantly increased the percentages of T lymphocytes and NK cells in peripheral blood,and regulated 15 immune-related cytokines were regulated in serum or colon or both,including immunoglobulin（Ig）,interleukin（IL）,Toll-like receptor（TLR）,interferon-γ（IFN-γ）,transforming growth factor-β（TGF-β）,triggering receptor expressed on myeloid cells-2,TREM-2)and granulocyte-macrophage colony-stimulating factor（GM-CSF）,causing the organism response develop to the direction of enhancing immunity and tumor inhibition.Meanwhile,ZQFZ up-regulated the levels of CD4,CD8 and IFN-γin spleen and colorectal tumors,and decreased cytotoxic T lymphocyte antigen 4（CTLA4）and programmed death-ligand 1（PD-L1）,confirming the activation of immune responses by ZQFZ in which cytotoxic T cell anticancer immune responses were enhanced,and enhanced the immunity of the organisms.In conclusion,the inhibitory effect of ZQFZ on CRC growth was demonstrated in Apc^Min/+mice,showing antitumor activity,and this effect may be achieved by modulating the abundance of specific gut microbes and increasing the immune response.This study provides an experimental basis for the application of ZQFZ as a therapeutic drug for CRC.